guideRNA和Cas9/Cpf1蛋白体外切割靶点DNA实验方法 化学合成crRNA+tracrRNA及Cas9蛋白,化学合成crRNA及 Cpf1蛋白(RNP)体外切割含靶点PCR产物实验。实验前需提前 PCR得到含靶点PCR产物(用DEPC水进行PCR)。 按以下表格混合各组分 1)加入5 μL PCR产物,混合后37℃反应1个小时; 2)加入1.0 μL(20 mg/mL)蛋白酶K,...
近日,《Nature Communications》杂志发表了一篇题为 Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes的文章,研究人员专为蚊子开发了一种Cas9/guide-RNA 表达“质粒工具包”,以遏制“病媒”蚊子传播病毒的能力。 https://doi.org/10.1038/s4146...
接下来,作者测试了通过这种RNA调节方法对CRISPR-Cas9切割活性的调节能力。在30分钟内,VE对sgRNA的修饰不会明显影响Cas9的切割活性,而在光照和添加PQ的条件下,Cas9的活性迅速下降;在类似的CRISPR-Cas13a系统中,VE和PQ也能高效地调节蛋白活...
aelectrification 电化[translate] arecombination–mediated genome 再结合斡旋的染色体[translate] aguide RNA and Cas9 指南RNA和Cas9[translate]
gRNA, or guide RNA, is part of the CRISPR-Cas9 system. The gRNA molecule has two components: a target-specific CRISPR RNA (crRNA) and an auxiliary trans-activating crRNA (tracrRNA). The gRNA guides the Cas9 protein to a specific genomic locus via base pairing between the 20-mer crRNA seq...
These data also revealed an age-related phenomenon in plants whereby the RNA suppressor P19 had more of an effect in older plants. Lastly, we used a single TRBO vector to express both Cas9 and a sgRNA. Taken together, we suggest that viral RNA suppressors could be used for further ...
pioneering evidence for an RNA virus vector, that the Cas9 protein and the sgRNA can be expressed from the same virus backbone, towards the implementation of single delivery platforms. Overall, it is evident that the integration of virus vector technology, suppressors, and CRISPR/Cas9 can be ...
在HeLa细胞中进行了全基因组CRISPR-Cas9筛选，GeCKO v2向导RNA ( gRNA )文库并进行3轮TcdB4选择 2. ...
CRISPR-Cas9基因组编辑技术是一种强大的工具,能够实现目标基因的调控。虽然这一技术能实现针对目标基因的高效编辑,但是同时也存在无法忽视的脱靶效应。在本文中,作者发展了一种基于RNA化学修饰以及光激活生物正交反应的针对sgRNA的修饰调控方法,实现了对细胞中CRISPR-Cas9基因编辑系统的光控终止以及脱靶效应的有效控制。
All-in-one sgRNA + Cas9 Stable guide RNA expression for cell types not amenable to transfection Lentiviral sgRNA Stable guide RNA expression for cell types not amenable to transfection Custom guide RNA solutions CRISPR Design Tool Design and order custom guide RNA for any species, nuclease or...